Unusual flexibility of distal and proximal histidine residues in the haem pocket of Drosophila melanogasterhaemoglobin

Abstract

Several pH-dependent low-spin ferric haem forms are identified in a frozen solution of the ferric ¹²¹Cys→Ser mutant of Drosophila melanogasterhaemoglobin (DmHb1*) using electron paramagnetic resonance (EPR) techniques. Different forms with EPR parameters typical of bis-histidine coordinated haem iron centers were observed. Strong pH-dependent changes in the EPR signatures were observed related to changes in the haem pocket. The pulsed EPR data indicate that both the distal and proximal histidine exhibit a large libration around the Fe–N_His axis. The resonance Raman spectra of the CO-ligated ferrous form of Drosophila melanogasterhaemoglobin are typical of an open conformation, with little stabilization of the CO ligand by the surrounding amino-acid residues. The EPR data of the cyanide-ligated ferric DmHb1* indicates a close similarity with cyanide-ligated ferric myoglobin. The structural characteristics of DmHb1* are found to clearly differ from those of other bis-histidine-coordinated globins.